The Role Of HSYA In The Proliferation,Migration And Chemoresistance To 5-FU Of Colorectal Cancer Cells

Background and Objective:Colorectal cancer（CRC）is one of the most common malignant tumors,and its incidence and mortality are increasing year by year.5-Fluorouracil（5-FU）is a first-line chemotherapeutic drug for CRC,but a considerable number of patients are resistant to 5-FU chemotherapy and cannot achieve ideal therapeutic effect.Therefore,it is of great significance to look for new anti-tumor active substances to ameliorate 5-FU chemotherapy resistance in patients with CRC.Hydroxysafflor yellow A（HSYA）,one of the important active components of traditional Chinese medicine safflower,has good anti-tumor activity.However,the effects of HSYA on the proliferation,migration and chemoresistance of CRC cells and its molecular mechanisms are rarely reported.The aim of this study is to elucidate the role of HSYA in the regulation of CRC cell proliferation,migration and 5-FU resistance and its related molecular mechanisms,and to provide new ideas for the clinical treatment of CRC.Methods:Firstly,the effect of HSYA on the proliferation of CRC cells was detected by MTT assay and colony formation assay.Secondly,the effect of HSYA on the cell cycle of CRC cells was detected by cell cycle assay.Furthermore,Wound healing assay and Transwell assay were used to detect the effect of HSYA on the migration of CRC cells.On this basis,DMSO treatment group,HSYA monotherapy group,5-FU monotherapy group and HSYA combined with 5-FU treatment group were set up to explore the effect of HSYA on 5-FU chemoresistance of CRC cells by MTT assay,colony formation assay and Transwell migration assay.Finally,western blot was used to analyze the effects of HSYA,5-FU and their combination on autophagy and AKT/mTOR signaling pathway in CRC cells.In addition,AKT activator SC79 was used to further verify whether HSYA could up-regulate the autophagy activity of CRC cells by inhibiting AKT/mTOR signaling pathway,thereby inhibiting the proliferation and migration of CRC cells and ameliorating their 5-FU chemotherapy resistance.Results:1.HSYA can significantly inhibit the proliferation and migration of CRC cells,and arrest the cell cycle in G₀/G₁ phase;2.HSYA can significantly enhance the inhibitory effect of 5-FU on the proliferation and migration of CRC cells.That is,HSYA can significantly ameliorate the 5-FU chemotherapy resistance of CRC cells;3.HSYA promotes 5-FU-induced autophagy in CRC cells through down-regulating AKT/mTOR signaling pathway;4.SC79 can reverse the promoting effect of HSYA on 5-FU-induced autophagy of CRC cells,and then reverse the ameliorating effect of HSYA on 5-FU chemoresistance of CRC cells.Conclusion:HSYA can upregulate the autophagy activity of CRC cells through inhibiting AKT/mTOR signaling pathway,thereby inhibiting the proliferation and migration of CRC cells and ameliorating the 5-FU chemotherapy resistance.HSYA is a promising candidate for the clinical treatment of CRC.